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Information on EC 6.3.2.3 - glutathione synthase and Organism(s) Escherichia coli and UniProt Accession P04425

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EC Tree
     6 Ligases
         6.3 Forming carbon-nitrogen bonds
             6.3.2 Acid—amino-acid ligases (peptide synthases)
                6.3.2.3 glutathione synthase
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This record set is specific for:
Escherichia coli
UNIPROT: P04425 not found.
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
glutathione synthetase, glutathione synthase, gsh synthetase, gshs, gsh-s, gsh synthase, tags2, gshii, gshs1, tags1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glutathione synthase
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-
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Glutathione synthetase
Glutathione synthetase (tripeptide)
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-
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GSH synthetase
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-
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GSHase
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-
-
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GSS
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-
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Phytochelatin synthetase
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-
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Synthetase, glutathione
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-
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione
show the reaction diagram
the catalytic mechanism is proposed to proceed via phosphorylation of the dipeptide substrate to yield an acyl phosphate intermediate. This intermediate is subsequently attacked by glycine, followed by loss of inorganic phosphate, leading to glutathione formation
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxamide formation
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carboxylic acid-amide formation
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SYSTEMATIC NAME
IUBMB Comments
gamma-L-glutamyl-L-cysteine:glycine ligase (ADP-forming)
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CAS REGISTRY NUMBER
COMMENTARY hide
9023-62-5
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
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-
-
?
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
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glutathione biosynthesis
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-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
UTP + gamma-Glu-L-Cys + Gly
UDP + phosphate + glutathione
show the reaction diagram
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10% of the activity relative to ATP
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-
?
additional information
?
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the loopless mutant shows gamma-Glu-L-Cys-dependent ATP hydrolase activity to almost the same extent as its glutathione synthetase activity
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-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + gamma-Glu-L-Cys + Gly
?
show the reaction diagram
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glutathione biosynthesis
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-
?
ATP + gamma-Glu-L-Cys + Gly
ADP + phosphate + glutathione
show the reaction diagram
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-
-
ir
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
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divalent metal ion required, Mg2+ is most effective, Co2+ can replace Mg2+
Mg2+
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divalent metal ion required, Mg2+ is most effective
Mn2+
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can replace Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoate)
7,8-dihydrofolate
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-
adenosine
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adenosine-5'-tetraphospho-5'-pyridoxal
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when incubated with an adenosine-5'-polyphospho-5'-pyridoxal or pyridoxal phosphate in the presence of Mg2+ and then reduced with sodium borohydride, it is most rapidly inactivated by adenosine-5'-tetraphospho-5'-pyridoxal, addition of either ATP or gamma-glutamylcysteine protects from inactivation
gamma-Glu-Cys
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above 2 mM
GSSG
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no inhibition by reduced glutathione
methotrexate
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NADPH
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weak
trimethoprim
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24 - 1.85
ATP
0.63 - 3.33
gamma-Glu-Cys
0.4 - 29.8
Gly
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Uniprot
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
152000
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gel filtration
38000
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x * 38000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 38000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure of Escherichia coli B glutathione synthetase complexed with ADP, glutathione, and sulfate at 2.0 A resolution
at 2.0 A resolution
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
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the crystal structure of the loopless mutant, in which the loop is replaced by 3 Gly residues, is identical with that of the wild-type enzyme
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under optimal catalytic condition pH 7.5
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G240V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
P227V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
P227V/G240V
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mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.5
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stable
1192
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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stable below
60
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15 min, 50% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
proteolysis by arginyl endopeptidase or trypsin causes a time-dependent decrease in activity. Only one peptide bond between Arg233 and Gly234 in the loop is cleaved
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relatively stable at all stages of purification
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several successive freezings and thawings of the cell-free extract have no effect on enzyme activity
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the remaining activity after arginyl-endopeptidase treatment is higher in the presence of ATP and/or gamma-Glu-Cys than in their absence
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 20% loss of activity after 2 months
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-20°C, stable for weeks
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloning and amplification of a gene for glutathione synthetase
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Escherichia coli B enzyme overproduced in Escherichia coli JM109
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Escherichia coli C600 cells transformed by a recombinant plasmid for the glutathione synthetase gene of Escherichia coli B
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mutants Cys122Ala, Cys195Ala, Cys222Ala and Cys289Ala show no critical loss of activity. Multiple replacement of Cys residues, however, decreases enzymatic activity to 45-26% of the activity of the wild-type enzyme
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overexpression in Brassica juncea plants via infection with Agrobacterium tumefaciens, under control of the 35S cauliflower mosaic virus promotor
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the gene for glutathione synthetase is polymerized and cloned onto the vector plasmid pBR325
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the gene for glutathione synthetase of Escherichia coli B is cloned onto vector plasmid pBR325. Escherichia coli C600 cells transformed by a recombinant plasmid pBR325
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kato, H.; Chihara, M.; Nishioka, T.; Murata, K.; Kimura, A.; Oda, J.
Homology of Escherichia coli B glutathione synthetase with dihydrofolate reductase in amino acid sequence and substrate binding site
J. Biochem.
101
207-215
1987
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Watanabe, K.; Yamano, Y.; Murata, K.; Kirmura, A.
Glutathione production by Escherichia coli cells with hybrid plasmid containing tandemly polymerized genes for glutathione synthetase
Appl. Microbiol. Biotechnol.
24
375-378
1986
Escherichia coli, Escherichia coli B / ATCC 11303
-
Manually annotated by BRENDA team
Gushima, H.; Miya, T.; Murata, K.; Kimura, A.
Purification and characterization of glutathione synthetase from Escherichia coli B
J. Appl. Biochem.
5
210-218
1983
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Murata, K.; Miya, T.; Gushima, H.; Kimura, A.
Cloning and amplification of a gene for glutathione synthetase in Escherichia coli B
Agric. Biol. Chem.
47
1381-1383
1983
Escherichia coli, Escherichia coli B / ATCC 11303
-
Manually annotated by BRENDA team
Apontoweil, P.; Berends, W.
Glutathione biosynthesis in Escherichia coli K12. Properties of the enzymes and regulation
Biochim. Biophys. Acta
399
1-9
1975
Escherichia coli
Manually annotated by BRENDA team
Meister, A.
Glutathione synthesis
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
10
671-697
1974
Saccharomyces cerevisiae, Escherichia coli, Homo sapiens, Pigeon
-
Manually annotated by BRENDA team
Wang, C.L.; Oliver, D.J.
Identification of a putative flexibel loop in Arabidopsis glutathione synthetase
Biochem. J.
322
241-244
1997
Arabidopsis sp., Escherichia coli
Manually annotated by BRENDA team
Tanaka, T.; Nishioka, T.; Oda, J.
Nicked multifunctional loop of glutathione synthetase still protects the catalytic intermediate
Arch. Biochem. Biophys.
339
151-156
1997
Escherichia coli
Manually annotated by BRENDA team
Hara, T.; Tanaka, T.; Kato, H.; Nishioka, T.; Oda, J.
Site-directed mutagenesis of glutathione synthetase from Escherichia coli B: mapping of the gamma-glutamyl-L-cysteine-binding site
Protein Eng.
8
711-716
1995
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Matsuda, K.; Mizuguchi, K.; Nishgioka, T.; Kato, H.; Go, N.; Oda, J.
Crystal structure of glutathione synthetase at optimal pH: domain architecture and structural similarity with other proteins
Protein Eng.
9
1083-1092
1996
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Fan, C.; Moews, P.C.; Shi, Y.; Walsh, C.T.; Knox, J.R.
A common fold for peptide synthetases cleaving ATP to ADP: glutathione synthetase and D-alanine:D-alanine ligase of Escherichia coli
Proc. Natl. Acad. Sci. USA
92
1172-1176
1995
Escherichia coli
Manually annotated by BRENDA team
Kato, H.; Tanaka, T.; Nishioka, T.; Kimura, A.; Oda, J.
Role of cysteine residues in glutathione synthetase from Escherichia coli B
J. Biol. Chem.
263
11646-11651
1988
Escherichia coli
Manually annotated by BRENDA team
Yamaguchi, H.; Kato, H.; Hata, Y.; Nishioka, T.; Kimura, A.; Oda, J.; Katsube, Y.
Three-dimensional structure of the glutathione synthetase from Escherichia coli B at 2.0 A resolution
J. Mol. Biol.
229
1083-1100
1993
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Hibi, T.; Kato, H.; Nishioka, T.; Oda, J.; Yamaguchi, H.; Katsube, Y.; Tanizawa, K.; Fukui, T.
Use of adenosine(5)poylphospho(5)pyridoxals to study the substrate-binding region of glutathione synthetase from Escherichia coli B
Biochemistry
32
1548-1554
1993
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Tanaka, T.; Yamaguchi, H.; Kato, H.; Nishioka, T.; Katsube, Y.; Oda, J.
Flexibility impaired by mutations related the multifunctional roles of the loop in glutathione synthetase
Biochemistry
32
12398-12404
1993
Escherichia coli
Manually annotated by BRENDA team
Kato, H.; Tanaka, T.; Yamaguchi, H.; Hara, T.; Nishioka, T.; Katsube, Y.; Oda, J.
Flexible loop that is novel catalytic machinery in a ligase. Atomic structure and function of the loopless glutathione synthetase
Biochemistry
33
4995-4999
1994
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Tanaka, T.; Kato, H.; Nishioka, T.; Oda, J.
Mutational and proteolytic studies on a flexible loop in glutathione synthetase from Escherichia coli B: the loop and arginine 233 are critical for the catalytic reaction
Biochemistry
31
2259-2265
1992
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Hara, T.; Kato, H.; Katsube, Y.; Oda, J.
A pseudo-Michaelis quarternary complex in the reverse reaction of a ligase: structure of Escherichia coli B glutathione synthetase complexed with ADP, glutathione, and sulfate at 2.0 A resolution
Biochemistry
35
11967-11974
1996
Escherichia coli (P04425), Escherichia coli
Manually annotated by BRENDA team
Zhu, Y.L.; Pilon-Smits, E.A.H.; Jouanin, L.; Terry, N.
Overexpression of glutathione synthetase in Indian mustard enhances cadmium accumulation and tolerance
Plant Physiol.
119
73-79
1999
Brassica juncea, Escherichia coli
Manually annotated by BRENDA team